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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Details EV-TRACK ID Experiment nr. Species Sample type Separation protocol First author Year EV-METRIC
EV120147 1/1 Acinetobacter baumannii Bacteria (d)(U)C
Filtration
UF
Moon DC 2012 11%

Study summary

Full title
All authors
Moon DC, Choi CH, Lee JH, Choi CW, Kim HY, Park JS, Kim SI, Lee JC
Journal
J Microbiol
Abstract
Acinetobacter baumannii secretes outer membrane vesicles (OMVs) during both in vitro and in vivo gro (show more...)Acinetobacter baumannii secretes outer membrane vesicles (OMVs) during both in vitro and in vivo growth, but the biogenesis mechanism by which A. baumannii produces OMVs remains undefined. Outer membrane protein A of A. baumannii (AbOmpA) is a major protein in the outer membrane and the C-terminus of AbOmpA interacts with diaminopimelate of peptidoglycan. This study investigated the role of AbOmpA in the biogenesis of A. baumannii OMVs. Quantitative and qualitative approaches were used to analyze OMV biogenesis in A. baumannii ATCC 19606T and an isogenic ?AbOmpA mutant. OMV production was significantly increased in the ?AbOmpA mutant compared to wild-type bacteria as demonstrated by quantitation of proteins and lipopolysaccharides (LPS) packaged in OMVs. LPS profiles prepared from OMVs from wild-type bacteria and the ?AbOmpA mutant had identical patterns, but proteomic analysis showed different protein constituents in OMVs from wild-type bacteria compared to the ?AbOmpA mutant. In conclusion, AbOmpA influences OMV biogenesis by controlling OMV production and protein composition. (hide)
EV-METRIC
11% (40th percentile of all experiments on the same sample type)
 Reported
 Not reported
 Not applicable
EV-enriched proteins
Protein analysis: analysis of three or more EV-enriched proteins
non EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody specifics
Protein analysis: antibody clone/reference number and dilution
lysate preparation
Protein analysis: lysis buffer composition
Study data
Sample type
Bacteria
Sample origin
NAY
Focus vesicles
OMV
Separation protocol
Separation protocol
  • Gives a short, non-chronological overview of the
    different steps of the separation protocol.
    • dUC = (Differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
(d)(U)C
Filtration
UF
Protein markers
EV: OmpA
non-EV:
Proteomics
yes
Show all info
Study aim
Biogenesis/Sorting
Sample
Species
Acinetobacter baumannii
Sample Type
Bacteria
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Equal to or above 150,000 g
Pelleting performed
Yes
Pelleting: time(min)
180
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Western Blot
Antibody details provided?
No
Detected EV-associated proteins
OmpA
ELISA
Antibody details provided?
No
Detected EV-associated proteins
OmpA
Characterization: Particle analysis
EM
EM-type
transmission EM
Image type
Wide-field
1 - 1 of 1
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV120147
species
Acinetobacter
baumannii
sample type
Bacteria
condition
NAY
separation protocol
(d)(U)C
Filtration
UF
Exp. nr.
1
EV-METRIC %
11